艾地苯醌对癫痫大鼠模型神经元线粒体功能、能量代谢及抗氧化作用机制研究  

Study on the Mechanism of Idebenone on Mitochondrial Function,Energy Metabolism and Antioxidation in Epileptic Rats Model

作　　者：崔小丽[1] 孙宁[1] 赵瑞 马妮[1] 贾瑞华 CUI Xiao-li;SUN Ning;ZHAO Rui;MA Ni;JIA Rui-hua(Department of Neurology,Shaanxi Provincial People’s Hospital,Xi’an 710068,China)

机构地区：[1]陕西省人民医院神经内一科,西安710068

出　　处：《现代检验医学杂志》2023年第1期44-48,65,共6页Journal of Modern Laboratory Medicine

基　　金：陕西省自然科学基础研究计划(2021JQ-903):艾地苯醌对癫痫持续状态大鼠学习记忆的影响及其机制的研究。

摘　　要：目的建立癫痫模型,研究艾地苯醌(Idebenone)的抗癫痫作用机制。方法30只SD大鼠,分为空白对照组(正常大鼠)、建模组和建模+艾地苯醌干预组(每组10只),建模组和干预组均采用经典氯化锂-匹罗卡品的癫痫诱导方法建立癫痫大鼠模型,艾地苯醌干预30天后检测大鼠血清、海马体组织中超氧化物歧化酶活性(superoxide dismutase,SOD)和丙二醛(malondialdehyde,MDA)含量的变化,通过Nissl染色评估艾地苯醌对神经元的保护作用。利用CCK-8明确艾地苯醌对癫痫细胞模型的最低适用浓度。在无镁诱导的癫痫细胞模型中检测ATP生成和线粒体膜电位,评价艾地苯醌对神经元线粒体产能功能的影响。结果建模组大鼠血清SOD活性为190.25±18.17 U/ml,较对照组(467.22±23.43 U/ml)降低,建模组海马组织中SOD活性(107.34±9.33 U/ml)较对照组(298.77±15.32 U/ml)降低,差异均具有统计学意义(t=-22.10,-16.30,均P<0.05);艾地苯醌干预后大鼠血清和海马组织中SOD活性有所恢复(384.79±29.21 U/ml,212.08±24.32 U/ml),与建模组相比升高,差异具有统计学意义(t=21.06,13.62,均P<0.05)。MDA的检测结果表明,与对照组相比(7.33±0.87 nmol/L),建模组大鼠血清中MDA含量增加(14.01±0.93nmol/L),海马组织中MDA含量(23.47±1.89 nmol/L)也较对照组(11.03±1.28 nmol/L)升高,差异具有统计学意义(t=5.72,9.19,均P<0.05);艾地苯醌干预后癫痫鼠血清与海马组织中MDA含量均下降(9.35±0.83 nmol/L,13.77±1.34 nmol/L),与造模组相比差异具有统计学意义(t=-17.68,-22.87,均P<0.05)。Nissl染色提示艾地苯醌干预后活性神经元数目增加(1977±200个/mm^(2)),与造模组(1387±146个/mm^(2))相比差异具有统计学意义(t=3.32,P<0.050)。细胞学实验表明,艾地苯醌干预能够提高线粒体ATP产能(0.92±0.14 vs 0.58±0.04),差异具有统计学意义(t=3.75,P=0.000);四甲基罗丹明甲酯(TMRM)荧光强度检测表明艾地苯醌干预后线粒体膜电位显著提高(0.97Objective To study the function of idebenone on neurons and the underlying mechanism in epileptic models.Methods 30 SD rats were divided into control group(normal rats),modeling group and modeling+idebenone intervention group(n=10 rats in each group).The classic lithium chloride-pilocarpine induced epilepsy model was established to detect the effects of idebenone on superoxide dismutase(SOD)activity and malondialdehyde(MDA)level in serum and hippocampus after 30 days of intervention.Function of idebenone on neurons was detected by Nissl staining.CCK-8 was used to determine the minimum appropriate concentration of idebenone for epilepsy cell models.ATP production and mitochondrial membrane potential were detected in the Mg-free epilepsy cell mode.Results The serum SOD activity in the modeling group was190.25±18.17 U/ml,which was lower than that in the control group(467.22±23.43 U/ml),and the SOD activity in the hippocampus of the modeling group(107.34±9.33 U/ml)was lower than that in the control group(298.77±15.32 U/ml).The differences were statistically significant(t=-22.10,-16.30,all P<0.05).SOD activity in serum and hippocampal tissue of rats recovered after intervention of idebenone,(384.79±29.21 U/ml,212.08±24.32 U/ml),which was higher than that of the modeling group,with statistical difference(t=21.06,13.62,all P<0.05).Compared with the control group(7.33±0.87nmol/L),the content of MDA in serum in the modeling group was increased(14.01±0.93 nmol/L).MDA content in hippocampus(23.47±1.89 nmol/L)was higher than that in control group(11.03±1.28 nmol/L),and the difference was statistically significant(t=5.72,9.19,all P<0.05).MDA in serum and hippocampal tissue of epileptic rats decreased(9.35±0.83nmol/L,13.77±1.34 nmol/L)after idebenone intervention,and the difference was statistically significant compared with the model group(t=-17.68,-22.87,P<0.05).Nissl staining indicated that the number of active neurons increased after idebenone intervention(1977±200 cells/mm^(2)),and the difference was statisti

关 键 词：癫痫 艾地苯醌 氧化应激 神经元 线粒体 

分 类 号：R446.19[医药卫生—诊断学]